Construction of THY1 eukaryotic expression plasmid and its effects on growth of ovarian cancer SKOV3 cells / 生物医学工程学杂志

ABSTRACT

This study was designed to construct THY1 eukaryotic expression plasmid and assess its effects on epithelial ovarian cancer cell line SKOV3. The gene fragment coding for THY1 was inserted into pcDNA3.1(+) for constructing the recombinant plasmid pcDNA3.1(+)-THY1. The eukaryotic expression plasmid was analyzed by PCR, restriction endonucleases digestion and DNA sequencing. The recombinant plasmid pcDNA3.1(+)-THY1 was transfected into SKOV3 cells by liposome protocol. The experimental cells were classified into three groups SKOV3-THY1, SKOV3-Null and SKOV3. The pcDNA3.1(+)-THY1 has been transfected into SKOV3 cells by RT-PCR and immunohistochemistry. The cell inhibitory rate of SKOV3-THY1 (56.6% at the fifth day) was higher than that of SKOV3-Null (12.5%), there was significant difference between them (P<0.05). The ratios of G1 phase of SKOV3 cells after transfection were increased and the ratios of S phase were decreased significantly. There was significant difference between SKOV3-THY1 and SKOV3-Null or SKOV3 (P<0.05), but there was no significant difference between SKOV3-Null and SKOV3 (P>0.05). We have constructed the recombinant plasmid pcDNA3.1(+)-THY1 sucessfully. THY1 transfection can inhibit the growth of SKOV3 cells in vitro.